PMID- 10436022
OWN - NLM
STAT- MEDLINE
DCOM- 19991012
LR  - 20181201
IS  - 1059-1524 (Print)
IS  - 1059-1524 (Linking)
VI  - 10
IP  - 8
DP  - 1999 Aug
TI  - Clathrin assembly lymphoid myeloid leukemia (CALM) protein: localization in
      endocytic-coated pits, interactions with clathrin, and the impact of
      overexpression on clathrin-mediated traffic.
PG  - 2687-702
AB  - The clathrin assembly lymphoid myeloid leukemia (CALM) gene encodes a putative
      homologue of the clathrin assembly synaptic protein AP180. Hence the biochemical 
      properties, the subcellular localization, and the role in endocytosis of a CALM
      protein were studied. In vitro binding and coimmunoprecipitation demonstrated
      that the clathrin heavy chain is the major binding partner of CALM. The bulk of
      cellular CALM was associated with the membrane fractions of the cell and
      localized to clathrin-coated areas of the plasma membrane. In the membrane
      fraction, CALM was present at near stoichiometric amounts relative to clathrin.
      To perform structure-function analysis of CALM, we engineered chimeric fusion
      proteins of CALM and its fragments with the green fluorescent protein (GFP).
      GFP-CALM was targeted to the plasma membrane-coated pits and also found
      colocalized with clathrin in the Golgi area. High levels of expression of
      GFP-CALM or its fragments with clathrin-binding activity inhibited the
      endocytosis of transferrin and epidermal growth factor receptors and altered the 
      steady-state distribution of the mannose-6-phosphate receptor in the cell. In
      addition, GFP-CALM overexpression caused the loss of clathrin accumulation in the
      trans-Golgi network area, whereas the localization of the clathrin adaptor
      protein complex 1 in the trans-Golgi network remained unaffected. The ability of 
      the GFP-tagged fragments of CALM to affect clathrin-mediated processes correlated
      with the targeting of the fragments to clathrin-coated areas and their
      clathrin-binding capacities. Clathrin-CALM interaction seems to be regulated by
      multiple contact interfaces. The C-terminal part of CALM binds clathrin heavy
      chain, although the full-length protein exhibited maximal ability for
      interaction. Altogether, the data suggest that CALM is an important component of 
      coated pit internalization machinery, possibly involved in the regulation of
      clathrin recruitment to the membrane and/or the formation of the coated pit.
FAU - Tebar, F
AU  - Tebar F
AD  - Department of Pharmacology, University of Colorado Health Science Center, Denver,
      Colorado 80111, USA.
FAU - Bohlander, S K
AU  - Bohlander SK
FAU - Sorkin, A
AU  - Sorkin A
LA  - eng
GR  - P30 CA046934/CA/NCI NIH HHS/United States
GR  - CA-46934/CA/NCI NIH HHS/United States
GR  - DK-46817/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Mol Biol Cell
JT  - Molecular biology of the cell
JID - 9201390
RN  - 0 (Adaptor Proteins, Vesicular Transport)
RN  - 0 (Clathrin)
RN  - 0 (Luminescent Proteins)
RN  - 0 (Monomeric Clathrin Assembly Proteins)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (PICALM protein, human)
RN  - 0 (Phosphoproteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (clathrin assembly protein AP180)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - EC 2.7.10.1 (ErbB Receptors)
SB  - IM
MH  - Adaptor Proteins, Vesicular Transport
MH  - Animals
MH  - Biological Transport
MH  - COS Cells/metabolism
MH  - Clathrin/genetics/*metabolism
MH  - Coated Pits, Cell-Membrane/metabolism
MH  - Endocytosis/*physiology
MH  - Endosomes/metabolism
MH  - ErbB Receptors/metabolism
MH  - Golgi Apparatus/metabolism
MH  - Green Fluorescent Proteins
MH  - Humans
MH  - Luminescent Proteins/genetics/metabolism
MH  - *Monomeric Clathrin Assembly Proteins
MH  - Nerve Tissue Proteins/genetics/*metabolism
MH  - Phosphoproteins/genetics/*metabolism
MH  - Recombinant Proteins/genetics/metabolism
PMC - PMC25500
EDAT- 1999/08/06 00:00
MHDA- 1999/08/06 00:01
CRDT- 1999/08/06 00:00
PHST- 1999/08/06 00:00 [pubmed]
PHST- 1999/08/06 00:01 [medline]
PHST- 1999/08/06 00:00 [entrez]
AID - 10.1091/mbc.10.8.2687 [doi]
PST - ppublish
SO  - Mol Biol Cell. 1999 Aug;10(8):2687-702. doi: 10.1091/mbc.10.8.2687.